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Fluoro Thiol ™ NEW!
Fluorescent Thiol Detection Kit - Detection of Thiols in tissue extracts /cell samples |
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| Introduction & Assay Principle | |||||||||
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During the normal course of metabolism, oxygen is partly reduced as electrons leak out of the electron transport chain during respiration. These partially reduced oxygen species (ROS) can react with organic substances through non-catalytic means. Furthermore, ROS can be generated via endogenous enzyme systems like plasma NADPH oxidase, cytoplasmic xanthine oxidase and organelle sources e.g., cytochrome P-450. ROS have been implicated in regulating diverse cellular functions including proliferation, defense against pathogens, intra-cellular signaling, transcriptional activation and apoptosis. Elevation of ROS beyond the buffering capacity of the cell can lead to oxidative stress. Elevated ROS levels can lead to damage of DNA/RNA, proteins and lipids which may lead to apoptosis. Cells have developed several mechanisms to counter act elevated ROS levels such as a thiol reducing buffer composed of cellular thiol levels (glutathione and thioredoxion) for the maintenance of the reduction-oxidation (redox) state of the cell, and enzymes to remove ROS (catalase, superoxide dismutase and glutathione peroxidase) (1-2).
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| Cell Technology’s Fluoro- Thiol kit detects Thiol levels in cells and tissue extracts. Hatsuo Maeda and co workers have developed 2,4-Dinitrobenzenesulfonyl fluorescein (dye) as a fluorescent specific probe for general thiol detection (3). The reaction scheme is outlined below.
Thiol (reduced) + non-fluorescent Dye -------> fluorescent analog
excitation: 488nm and emission at 515-530 nm
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1. Gamaley IA and Klyubin IV (1999) Roles of reactive oxygen species: Signaling and regulation of cellular functions. Int Rev Cytol 188:203–238..
2. Nakamura H, Nakamura K and Yodoi J (1997) Redox regulation of cellular activa-tion. Annu Rev Immunol 15:351–369.
3. 2,4-Dinitrobenzenesulfonyl Fluoresceins as Fluorescent Alternatives to Ellman s Reagent in Thiol-Quantification Enzyme Assays*. Hatsuo Maeda,*Hiromi Matsuno,Mai Ushida, Kohei Katayama,Kanako Saeki,and Norio Itoh. Angew.Chem.Int.Ed.2005 ,44 ,2922 –2925
4. Marchetti,P.,et al .,Redox regulation of apoptosis: impact of thiol oxidation status on mitochondrial function.Eur.J.Immunol.,27 ,289-296 (1997).
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| Kit contents (for 100 assays) | |||||||||
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Fluoro Thiol ™ Fluorescent Thiol Detection Kit - Detection of Thiols in tissue extracts /cell samples/荧光硫醇检测试剂盒---- 检测组织提取物或者细胞样品中的硫醇
主要优点：
检测组织提取物或者细胞样品中的硫醇。
灵敏的荧光检测
检测细胞凋亡，新陈代谢，氧化应激中减少的硫醇
检测细菌，真菌，植物细胞或者其他样品中减少的硫醇
96孔荧光酶标仪（激发波长：488nm，发射波长：515-530nm）
介绍和检测原理：
在正常的新陈代谢过程中，氧气会在代谢过程中随着电子传递链有一部分的减少。这部分减少的活性氧（ROS）会通过非催化方法与有机物质反应。此外，ROS可以通过内源性酶系统如NADPH氧化酶，细胞质黄嘌呤氧化酶以及细胞器如细胞色素P – 450而产生。 ROS可以参与调控多种细胞内的功能，包括增殖，抵御病原体的干扰，细胞内的信号转导，转录激活和细胞凋亡。 ROS的水平如果超出了细胞的缓冲能力将会导致氧化应激。 ROS水平升高会导致DNA / RNA，蛋白质和脂质的损伤，以及导致细胞凋亡。细胞有多种机制用来抵抗ROS水平的升高，如细胞内硫醇水平的降低能够维持细胞内氧化还原水平，以及去除活性氧的酶（过氧化氢酶，超氧化物歧化酶和谷胱甘肽过氧化物酶）（1-2）。
反应：
Cell Technology荧光硫醇检测试剂盒用来检测组织提取物和细胞样品中的硫醇水平。 Hatsuo Maeda和他的工作人员一起研发了一种荧光染料作为一种检测硫醇的荧光探针(3). 反应如下：
硫醇 (减少的) +非荧光染料------->荧光类似物
激发波长: 488nm 发射波长:515-530 nm
